ABSTRACT
Objective:To investigate the inhibitory effect of miR-497 on the corneal epithelial healing in diabetic mice and its possible mechanism.Methods:Forty healthy clean-grade wild-type C57BL/J6 mice were randomly divided into a blank control group and a model control group, with 20 mice in each group.Another 20 CRISPR/Cas9-mediated miR-497 knockout mice and miR-497 overexpression mice were taken as miR-497 knockout and miR-497 overexpression groups, respectively.The diabetes model was constructed by continuous intraperitoneal injection of streptozotocin (STZ) to the mice in model control, miR-497 knockout and miR-497 overexpression groups, and the mice in blank control group were injected with an equal amount of citrate buffer, followed by 8-week normal feeding.After the establishment of diabetes model, the corneal epithelial injury model was further constructed by scraping off part of the corneal epithelium with a central diameter of 2 mm.The corneal epithelial defect area of mice in 0, 12, 24 and 36 hours after corneal epithelial injury was observed by corneal fluorescein sodium staining.The expression of Wnt3a and β-catenin proteins in mice corneal tissues was detected by Western blot.The expression of miR-497 as well as the mRNA expression levels of cell proliferation-associated factor genes CyclinD1, c-Myc, and Ki-67 mRNA was detected by real-time quantitative fluorescence PCR.The targeting relationship between miR-497 and wnt3a was detected by a dual luciferase reporter gene assay.Human corneal epithelial cells (HCEC) were cultured in vitro and transfected with miR-497 mimics, miR-497 mimics negative control, miR-497 inhibitor, and miR-497 inhibitor negative control by Lipo8000 as miR-497 mimics group, mimics negative control group, miR-497 inhibitor group, andmiR-497 inhibitor negative control group, respectively, all of which were cultured in high glucose medium containing 25% glucose.Another two groups of HCEC were taken and cultured in medium containing 5% and 25% glucose as control and high glucose groups, respectively.The cell proliferation viability was determined by CCK8 method.The use and care of animals complied ith the ARVO statement.The study protocol was approved by the Ethics Committee of Renmin Hospital of Wuhan University (2019K-K010). Results:Eight weeks after STZ injection, the blood glucose of mice was significantly higher and the weight was significantly lower in each diabetic model group than those of blank control group (all at P<0.05). At 12, 24 and 36 hours after the corneal epithelial injury, the percentages of corneal epithelial defect area observed by slit-lamp microscopy in model control group were significantly higher than those in blank control group and miR-497 knockout group and lower than those in miR-497 overexpression group, and the differences were statistically significant (all at P<0.05). The relative expressions of wnt3a and β-catenin proteins in the corneal tissues of model control group were significantly lower than those of blank control group and miR-497 knockout group, but higher than those of miR-497 overexpression group, and the differences were statistically significant (all at P<0.05). The relative expressions of CyclinD1, c-Myc and Ki-67 mRNA in model control group were lower than those in miR-497 knockout group, but higher than those in miR-497 overexpression group, and the differences were statistically significant (all at P<0.05). The relative expression of miR-497 in model control group, miR-497 knockout group and miR-497 overexpression group was 1.00±0.02, 0.63±0.06 and 1.48±0.03, respectively, with a statistically significant difference ( F=19.62, P<0.01). The luciferase activity of miR-497-5p mimics group in wild-type wnt3a transfected cells was lower than that of miR-497-5p negative control group and empty vector group, and the differences were statistically significant (all at P<0.05). In the mutant wnt3a transfected cells, there was no significant difference in the luciferase activity among various groups ( F=0.73, P=0.59). The cell proliferation A value of high glucose group was 0.59±0.03, which was significantly lower than 0.59±0.03 of normal control group and 0.88±0.08 of miR-497 inhibitor group, but significantly higher than 0.48±0.11 of miR-497 mimics group (all at P<0.05). Conclusions:The silencing of miR-497 may promote the repair of diabetic corneal epithelial defects by targeting wnt/β-catenin pathway.
ABSTRACT
Objective:To investigate the role of microRNA (miR)-497 in the formation of corneal neovascularization (CNV) induced by alkali burn and its mechanism.Methods:Forty-two wild type (WT) C57BL/6 mice aged 6 to 8 weeks, 42 CRISPR/Cas9 mediated miR-497 knockout (KO) and 42 CRISPR/Cas9 mediated overexpression transgenic (TG) C57BL/6 mice were selected and assigned as WT group, KO group and TG group, respectively.The corneal alkali burn model was established.At 3, 7, 14 and 21 days after modeling, corneal epithelium damage and stromal turbidity were scored according to slit lamp microscopy.The area of neovascularization was measured.Corneal structural changes and expression of inflammatory cells were observed by histopathological staining.The expression of CD31 in corneal tissues was detected by immunohistochemistry staining.The targeted binding relationship between miR-497 and signal transducer and activator of transcription 3 (STAT3) was detected by luciferase reporter assay.The relative expressions of miR-497, vascular endothelial growth factor A (VEGFA), tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β and macrophage inflammatory protein (MCP)-1 mRNA were detected by real-time quantitative PCR.At 14 days following modeling, the expression of STAT3 and p-STAT3 proteins in mice corneal tissues was detected by Western blot.The use and care of animals complied with the ARVO statement.The study protocol was approved by the Ethics Committee of Renmin Hospital of Wuhan University (No.2019K-K010).Results:Corneal injury, inflammatory cell infiltration and CNV occurred in mice cornea after alkali burn.Corneal epithelial injury score, corneal stromal turbidity score and CNV area increased first and reached the peak on the 14th day after modeling, and then decreased.There were significant differences in corneal epithelial injury score, corneal stromal turbidity score, CNV area and number of CD31-positive cells among various time points after alkali burn ( Fgroup=49.19, 34.56, 44.56, 77.56; all at P<0.01; Ftime=51.62, 65.62, 71.32, 46.12; all at P<0.01). Corneal epithelial injury score, corneal stromal turbidity score, CNV area and the number of CD31-positive cells were greater in KO group at various time points than in WT and TG groups, and those in WT group were greater than in TG group (all at P<0.05). In WT STAT3 co-transfected cells, the luciferase activity of the miR-497 group was significantly lower than that of the miR-negative control group and normal control group (both at P<0.05). In mutant STAT3-transfected cells, there was no significant difference in luciferase activity among all groups ( F=0.69, P=0.56). On the 14th day after modeling, the relative expression levels of miR-497 in corneal tissue of WT, KO and TG groups were 0.68±0.11, 0.41±0.06 and 1.05±0.14, respectively, which were significantly lower than 1.00±0.04, 0.56±0.07 and 1.34±0.11 before modeling (all at P<0.01). The relative expressions of STAT3 and p-STAT3 were higher in KO group than in WT and TG groups, and were lower in TG group than in WT group, and the differences were statistically significant (all at P<0.05). The expressions of VEGFA, TNF-α, IL-6, IL-1β and MCP-1 mRNA at various time points after modeling in various groups were significantly higher than before modeling, which were higher in KO group than in WT and TG groups and were lower in TG group than in WT group, and the differences were statistically significant (all at P<0.01). Conclusions:MiR-497 inhibits corneal inflammation and CNV formation induced by alkali burn.It might inhibit the activation of the inflammation signal pathway via targeting STAT3.
ABSTRACT
With the improvement of computing power, the refinement of learning algorithms and architectures and the disclosure of big data, many achievements pertaining to artificial intelligence (AI) have been achieved in the field of medicine.In the field of ocular anterior segment, AI can provide adjunctive diagnosis for pterygium, corneal injury and fungal keratitis etc.In addition, AI presents potential value in screening, early diagnosis and management of keratoconus.For cataract, AI is expected to aid telemedicine and optimize IOL calculation.AI also shows good performance in diagnosing narrow anterior chamber angle, identifying the mechanism of angle closure, assessing the image quality of anterior chamber angle, and predicting the efficacy of glaucoma surgery.In optometry, AI appears to be available in predicting myopia progress, indicating risk factors and subjects of high myopia, and providing help for selection of orthokeratology and cornea refractive surgery.The development, application and limitation of AI technology in the field of ocular anterior segment and myopia management were reviewed.
ABSTRACT
Objective:To explore the clinical value of a diagnostic system of ophthalmic B-scan ultrasound images based on deep convolutional neural network.Methods:A total of 3 600 ophthalmic B-scan ultrasound images of 1 278 patients with an average age of (49.32±7.69) years at the Eye Center of Renmin Hospital of Wuhan University from January 2018 to October 2020 were collected to build an image database.These B-scan images were labeled by three ophthalmologists.The database was divided into the training dataset of 2 812 images and the testing dataset of 788 images.The deep learning algorithm was used to build a diagnostic model, which can identify retinal detachment (RD), vitreous hemorrhage (VH) and posterior vitreous detachment (PVD), and the accuracy of the model was evaluated.Another 120 B-scan ultrasound images were collected for the human-computer comparison between the model and 3 senior ophthalmologists.Eight junior clinicians were selected to evaluate another 150 B-scan images with and without the assistance of the model, and the accuracy was analyzed to evaluate the effect of the model.This study adhered to the Declaration of Helsinki and the study protocol was approved by Renmin Hospital of Wuhan University (No.WDRY2020K-192).Results:The accuracy of the model for identifying normal fundus, RD, VH, PVD and other diseases were 0.954, 0.909, 0.881, 0.990 and 0.920, respectively.The accuracy of the model was similar to that of senior doctors, and the time the model used was almost half that of doctors.With the assistance of the model, the diagnostic accuracy of the 8 junior clinicians who participated in the training was significantly improved ( P<0.01). Conclusions:The accuracy of RD, VH and PVD identification of the intelligent evaluation system is good, and the system can improve the accuracy and efficiency of clinical examinations, and can better assist clinicians in clinical evaluation.
ABSTRACT
As the frontline health care workers at the center of the novel coronavirus disease (COVIN-19) outbreak, we have found many asymptomatic COVIN-19 patients or patients with mild symptoms since December 2019. A number of COVIN-19 cases with conjunctivitis or conjunctivitis as the first symptom have been observed in our clinical work. This paper reports the diagnosis and treatment of one COVIN-19 patient with conjunctivitis as the first symptom and one COVIN-19 patient with conjunctivitis. Case one occurred conjunctivitis at the third day after closely contacted with determined COVID-19 patient and visited to eye doctor, and the symptom of conjunctivitis following the topical administration of anti-virual eyedrops for 1 week, followed by COVID-19. Her 2019-nCoV RNA detection of nasopharynx swab was positive but that of conjunctival sac swab was a negative result. Case two had a positive epidemiological history and simultaneous onset of COVID-19 and conjunctivitis.She presented positive results of 2019-nCoV RNA detection in both nasopharynx and conjunctival sac swabs, and other lab results supported the diagnosis of COVID-19 but she had a normal CT findings of the chest. The ocular symptoms were disappeared after topical administration of anti-virual eyedrops for 1 week.
ABSTRACT
At the end of 2019, the coronavirus disease-19 (COVID-19) broke out in Wuhan and has spread throughout the world, bringing great challengees to the public health, prevention, and control work in our country and globally.Although the national medical staff and Public Health Epidemic Prevention personnel are fighting the epidemic, the large numbers of patients and the long-term isolation measures greatly impacted people's lifestyles.The epidemic's development has currently been eased, but the blockades and quarantines in many cities have not been completely lifted.While controlling the coronavirus, lifestyle changes caused by long-term isolation, as well as fear of the disease and economic recession have brought people various negative emotions.These mental health issues may cause or aggravate the occurrence and development of dry eye.Therefore, the authors explore the impact of mental health on dry eye during the COVID-19 epidemic, alongside possible mechanisms, and put forward relevant suggestions for managing eye-related diseases during and after the epidemic.
ABSTRACT
As the frontline health care workers at the center of the novel coronavirus disease (COVID-19) outbreak, we have encountered many asymptomatic COVID-19 patients or patients with mild symptoms since December 2019.A number of COVID-19 cases with conjunctivitis or conjunctivitis as the first symptom have been observed in our clinical work.This paper reports the diagnosis and treatment of one COVID-19 patient with conjunctivitis as the first symptom and one COVID-19 patient with conjunctivitis.In case one conjunctivitis occurred at the third day after patient came in close contact with determined COVID-19 patient and visited an eye doctor, and the symptom of conjunctivitis following the topical administration of anti-viral eyedrops for 1 week, followed by COVID-19.Her 2019-nCoV RNA detection of nasopharynx swab was positive but that of conjunctival sac swab was a negative result.Case two had a positive epidemiological history and simultaneous onset of COVID-19 and conjunctivitis.She presented positive results of 2019-nCoV RNA detection in both nasopharynx and conjunctival sac swabs, and other lab results supported the diagnosis of COVID-19 but she had a normal CT findings of the chest.The ocular symptoms were disappeared after topical administration of anti-viral eyedrops for 1 week.
ABSTRACT
Objective:To investigate the effects of self-made Hezhong-Jianpi Decoction on gastrointestinal motility and level of inflammatory cell factors in patients with reflux esophagitis. Methods:A total of 102 patients with reflux esophagitis who met the inclusion criteria in our hospital from May 2016 to September 2018 were divided into two group according to the random number table method, 51 cases in each group. The control group was treated with conventional western medicine. The intervention group was treated with self-made Hezhong-Jianpi Decoction based on the control group. Both groups were treated for 8 weeks. The levels of serum gastrin (GAS) and motilin (MTL) were detected by radioimmunoassay, the levels of TNF-α, IL-6 and IL-17 were detected by ELISA, the adverse reactions during the treatment were recorded, and the clinical efficacy was evaluated. Results:The total effective rate of the experimental group was 92.1% (47/51) and that of the control group was 72.5% (37/51). The difference between the two groups was statistically significant ( χ2=6.746, P=0.009). After the treatment, the levels of serum GAS (152.93 ± 26.85 ng/L vs. 134.30 ± 29.88 ng/L, t=3.312), MTL (291.58 ± 29.75 ng/L vs. 262.40 ± 38.71 ng/L, t=4.268) in the intervention group were significantly higher than those in the control group ( P<0.05), and serum TNF-α, IL-6, IL-17 levels were significantly lower than those in the control group ( t value were 4.653, 5.362, 4.759, respectively, all Ps<0.01). During the treatment, the incidence of adverse reactions was 9.8% (5/51) in the control group and 12.0% (6/51) in the intervention group, and there was no significant difference between the two groups ( χ2=0.102, P=0.750). Conclusions:The self-made Hezhong-Jianpi Decoction combined with the conventional western medicine can promote the gastrointestinal motility of patients with reflux esophagitis, reduce the inflammatory reaction and improve the clinical efficacy.
ABSTRACT
Objective To compare the etiological constitution of recurrent miscarriage (RM) between patients with consecutive two and three or more miscarriages through combining the routine examination results and embryonic karyotype. Methods Patients with a history of two or more consecutive clinical miscarriages(≤12 weeks of gestation)consulting in the RM clinic of the First Affiliated Hospital of Sun Yat-sen University from March 2011 to January 2016 were collected. Six hundred and ninety-six with detailed history recorded, routine clinical examinations of RM and at least once embryonic karyotype were ultimately enrolled in this study. Their etiological constitution of RM were analyzed in groups of consecutive two and three or more miscarriage. The etiologies of RM in analysis consisted of women age, body mass index (BMI), chromosome abnormalities of couples, uterine abnormalities, endocrinology abnormalities and antiphospholipid syndrome(APS). Results (1)Among 696 patients, the abnormal embryonic karyotypes was 60.6%(422/696)and routine RM etiologies was 32.2%(224/696), leaving the ratio of unexplained RM was only 29.0%(202/696).(2)A total of 717 embryo karyotype were found in 696 patients, included 21 cases with twice embryo karyotype results the percentage of normal embryo was 39.7%(285/717), while abnormal ones was 60.3%(432/717). Among the types of abnormal karyotype, the most common ones (>10%)were trisomy 16(19.2%, 83/432), monosome X(11.3%, 49/432)and trisomy 22(10.9%, 47/432). (3)Among the 696 RM patients, the number of two and three or more miscarriages were respectively 446(64.1%,446/696)and 250(35.9%,250/696). Comparing groups of three or more miscarriages with two miscarriages, there were significant differencein older age as well as uterine adhesion(P<0.05). But no difference was found in body mass index(BMI), the rates of chromosome abnormalities of couples, uterine abnormalities except uterine adhesion, endocrinology abnormalities and APS (all P>0.05) between two groups. Conclusions The abnormal embryonic karyotype is the most common cause of first-trimester RM. The etiological constitution of two and three or more recurrent miscarriages is accordant, suggesting that routine clinical examination and the embryonic karyotype should be started following two consecutive clinical early miscarriages.
ABSTRACT
Dry eye is a common ocular disease with a high morbidity.Many factors contribute to dry eye with cause pathophysiological changes of the ocular surface.In recent years,with the increasing attention of dry eye and the research progress,people have more in-depth understanding of the dry eye.This article reviews the researches in recent years and new progress in the treatment of dry eye.
ABSTRACT
The corneal epithelial cells are at the surface of the cornea.Human corneal epithelial cells cultured in vitro contribute to fully understand the biological properties of corneal cells,construct disease models in vitro,and play its clinical significance better.This article reviews the recent advances in human corneal epithelial cells cultured in vitro about extractive fraction,extraction methods,factors of progress,impact identification methods.